Insights Into Rapid Screening and Activity Investigation of Novel Hemp Seed Pancreatic Lipase/Cholesterol Esterase Inhibitory Peptides

YIN Hao; ZHU Jiangxiong; ZHONG Yu; WANG Danfeng; DENG Yun; ZHANG Minyan; ZHAO Fang

doi:10.13386/j.issn1002-0306.2024050168

Turn off MathJax

Article Contents

Abstract

References

YIN Hao, ZHU Jiangxiong, ZHONG Yu, et al. Insights Into Rapid Screening and Activity Investigation of Novel Hemp Seed Pancreatic Lipase/Cholesterol Esterase Inhibitory Peptides[J]. Science and Technology of Food Industry, 2025, 46(9): 1−11. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024050168.

Citation:

PDF (4762 KB)

Insights Into Rapid Screening and Activity Investigation of Novel Hemp Seed Pancreatic Lipase/Cholesterol Esterase Inhibitory Peptides

1.
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China
2.
Yunnan Dali Research Institute of Shanghai Jiao Tong University, Dali 671000, China
3.
Eryuan County Inspection and Testing Institute, Dali Bai Autonomous Prefecture, Eryuan 671208, China

More Information

Received Date: May 15, 2024
Available Online: March 06, 2025

Graphical Abstract

Abstract

Abstract

This study aims to rapidly screen and identify pancreatic lipase (PL) and cholesterol esterase (CE) inhibitory peptides from hemp seed protein hydrolysates (HSPHs), and validate their activities, accompanied by preliminarily analyzing their molecular mechanisms. Initially, the PL and CE inhibitory activities of HSPHs were explored under different buffer systems. Subsequently, a combination of ultrafiltration, peptidomics, and virtual screening methods was employed for the rapid identification and screening of PL and CE inhibitory peptides. Finally, their activities were validated, and molecular docking was used to investigate the types of molecular interactions involved. The results indicated that, compared to conventional buffer systems, the simulated intestinal fluid system reduced the inhibitory effect of HSPHs on PL but enhanced their inhibitory effect on CE. Therefore, subsequent tests of PL and CE inhibitory activities were conducted using the simulated intestinal fluid system. The fractions of HSPHs smaller than 3 kDa, released by alcalase and papain, exhibited the highest PL/CE inhibitory activities. After identifying the peptide sequences, 136 peptides with higher content and potential activity were selected to establish a candidate peptide library. From this library, 14 peptides with lower binding energies were identified as PL and CE inhibitory peptides. In vitro activity validation demonstrated that APAM and RLPA exhibited significant inhibitory effects on both PL and CE. Molecular docking results suggested that APAM and RLPA might inhibit the activities of PL and CE by forming hydrogen bonds and hydrophobic interactions with the residues at the enzyme active sites and surface loops, thereby restricting the exposure of the active sites and the entry of substrates. This study indicates that hemp seeds are a potential source for developing lipid digestion enzyme inhibitory peptides, providing a reference for the subsequent development of lipid-lowering peptide products from hemp seeds.
- hemp seed,
- bioactive peptide,
- enzyme inhibiting,
- peptidomics,
- virtual screening,
- molecular docking

FullText(HTML)

References (42)

References

[1]	吕知新, 任龙梅, 李银换. 新时期工业大麻产业发展的机遇与挑战[J]. 南方农机,2021,52(23):117−119. [LÜ Z X, REN L M, LI Y H. Opportunities and challenges in the development of the industrial hemp industry in the new era[J]. China Southern Agricultural Machinery,2021,52(23):117−119.] doi: 10.3969/j.issn.1672-3872.2021.23.037 LÜ Z X, REN L M, LI Y H. Opportunities and challenges in the development of the industrial hemp industry in the new era[J]. China Southern Agricultural Machinery, 2021, 52（23）: 117−119. doi: 10.3969/j.issn.1672-3872.2021.23.037
[2]	何锦风, 陈天鹏, 卢蓉蓉, 等. 汉麻籽的综合利用及产业化研究[J]. 中国食品学报,2010,10(3):98−112. [HE J F, CHEN T P, LU R R, et al. Study on the comprehensive utilization and industrialization of hempseed (Fructus cannabis)[J]. Journal of Chinese Institute of Food Science and Technology,2010,10(3):98−112.] doi: 10.3969/j.issn.1009-7848.2010.03.016 HE J F, CHEN T P, LU R R, et al. Study on the comprehensive utilization and industrialization of hempseed （Fructus cannabis）[J]. Journal of Chinese Institute of Food Science and Technology, 2010, 10（3）: 98−112. doi: 10.3969/j.issn.1009-7848.2010.03.016
[3]	孟妍, 曾剑华, 王尚杰, 等. 汉麻籽蛋白研究进展[J]. 食品工业,2020,41(1):268−273. [MENG Y, ZENG J H, WANG S J, et al. Research progress on hemp seed protein[J]. The Food Industry,2020,41(1):268−273.] MENG Y, ZENG J H, WANG S J, et al. Research progress on hemp seed protein[J]. The Food Industry, 2020, 41（1）: 268−273.
[4]	GILDA A, CARMEN L, GIOVANNA B, et al. Exploration of potentially bioactive peptides generated from the enzymatic hydrolysis of hempseed proteins[J]. Journal of Agricultural and Food Chemistry,2017,65(47):10174−10184. doi: 10.1021/acs.jafc.7b03590
[5]	魏连会, 宋淑敏, 董艳, 等. 火麻籽多肽对高脂饮食喂养大鼠血脂的影响[J]. 食品科学,2021,42(11):161−167. [WEI L H, SONG S M, DONG Y, et al. Effect of hemp seed peptide on blood lipids in high-fat diet fed rats[J]. Food Science,2021,42(11):161−167.] doi: 10.7506/spkx1002-6630-20200421-266 WEI L H, SONG S M, DONG Y, et al. Effect of hemp seed peptide on blood lipids in high-fat diet fed rats[J]. Food Science, 2021, 42（11）: 161−167. doi: 10.7506/spkx1002-6630-20200421-266
[6]	World Health Organization. World obesity atlas 2023[R]. Geneva:WHO,2024.
[7]	NCD Risk Factor Collaboration. Worldwide trends in underweight and obesity from 1990 to 2022:A pooled analysis of 3663 population-representative studies with 222 million children, adolescents, and adults[J]. Lancet,2024,403(10431):1027−1050. doi: 10.1016/S0140-6736(23)02750-2
[8]	LIU T, LIU X, CHEN Q, et al. Lipase inhibitors for obesity:A review[J]. Biomedicine and Pharmacotherapy,2020,128:110314. doi: 10.1016/j.biopha.2020.110314
[9]	MUDGIL P, BABA W N, KAMAL H, et al. A comparative investigation into novel cholesterol esterase and pancreatic lipase inhibitory peptides from cow and camel casein hydrolysates generated upon enzymatic hydrolysis and in-vitro digestion[J]. Food Chemistry,2022,367:130661. doi: 10.1016/j.foodchem.2021.130661
[10]	KUMAR A, CHAUHAN S. Pancreatic lipase inhibitors:The road voyaged and successes[J]. Life Sciences,2021,271:119115. doi: 10.1016/j.lfs.2021.119115
[11]	ESFANDI R, SEIDU I, WILLMORE W, et al. Antioxidant, pancreatic lipase, and α-amylase inhibitory properties of oat bran hydrolyzed proteins and peptides[J]. Journal of Food Biochemistry,2022,46:e13762.
[12]	相欢, 崔春. 沙棘籽粕蛋白肽的稳定性及分离纯化[J]. 食品科学,2023,44(18):49−57. [XIANG H, CUI C. Stability and separation of peptides from seabuckthorn seed protein[J]. Food Science,2023,44(18):49−57.] XIANG H, CUI C. Stability and separation of peptides from seabuckthorn seed protein[J]. Food Science, 2023, 44（18）: 49−57.
[13]	钟婉滢, 苗建银, 叶灏铎, 等. 藜麦蛋白肽的酶解制备及体外降血脂与降尿酸活性研究[J]. 食品工业科技,2023,44(23):156−166. [ZHONG W Y, MIAO J Y, YE H D, et al. Enzymatic preparation of quinoa protein peptides and its lipid lowering and uric acid-lowering activity in vitro[J]. Science and Technology of Food Industry,2023,44(23):156−166.] ZHONG W Y, MIAO J Y, YE H D, et al. Enzymatic preparation of quinoa protein peptides and its lipid lowering and uric acid-lowering activity in vitro[J]. Science and Technology of Food Industry, 2023, 44（23）: 156−166.
[14]	ZHANG Y, TANG X, LI F, et al. Inhibitory effects of oat peptides on lipolysis:A physicochemical perspective[J]. Food Chemistry,2022,396:133621. doi: 10.1016/j.foodchem.2022.133621
[15]	YOU H, ZHANG Y, WU T, et al. Identification of dipeptidyl peptidase IV inhibitory peptides from rapeseed proteins[J]. LWT,2022,160:113255. doi: 10.1016/j.lwt.2022.113255
[16]	张李君, 赵甜甜, 陈杰琼, 等. 鲟鱼子酶解产物对酒精损伤肝细胞的保护作用及活性肽虚拟筛选[J]. 食品工业科技,2024,45(19):316−324. [ZHANG L J, ZHAO T T, CHEN J Q, et al. Protective effects of enzymatic products from sturgeon roe on alcohol-induced hepatic cell damage and virtual screening of active peptides[J]. Science and Technology of Food Industry,2024,45(19):316−324.] ZHANG L J, ZHAO T T, CHEN J Q, et al. Protective effects of enzymatic products from sturgeon roe on alcohol-induced hepatic cell damage and virtual screening of active peptides[J]. Science and Technology of Food Industry, 2024, 45（19）: 316−324.
[17]	刘芬娣, 李婧铭, 陈洪娇, 等. 生姜蛋白酶水解产物中二肽基肽酶-Ⅳ抑制肽的虚拟筛选及活性分析[J]. 现代食品科技,2024,40(5):34−42. [LIU F D, LI J M, CHEN H J, et al. Virtual screening and activity evaluation of dipeptidyl peptidase-IV inhibitory peptides from ginger protease hydrolysates[J]. Modern Food Science and Technology,2024,40(5):34−42.] LIU F D, LI J M, CHEN H J, et al. Virtual screening and activity evaluation of dipeptidyl peptidase-IV inhibitory peptides from ginger protease hydrolysates[J]. Modern Food Science and Technology, 2024, 40（5）: 34−42.
[18]	宋炜昱, 尹浩, 钟宇, 等. 不同品种汉麻籽蛋白质结构与功能特性分析[J]. 食品工业科技,2023,44(10):47−53. [SONG W Y, YIN H, ZHONG Y, et al. Analysis of protein structure and functional properties of hemp seeds of different varieties[J]. Science and Technology of Food Industry,2023,44(10):47−53.] SONG W Y, YIN H, ZHONG Y, et al. Analysis of protein structure and functional properties of hemp seeds of different varieties[J]. Science and Technology of Food Industry, 2023, 44（10）: 47−53.
[19]	YIN H, JIANG Y, ZHOU X, et al. Effect of radio frequency, ultrasound, microwave-assisted papain, and alcalase hydrolysis on the structure, antioxidant activity, and peptidomic profile of Rosa roxburghii Tratt. seed protein[J]. Journal of food science,2022,87(9):4040−4055. doi: 10.1111/1750-3841.16266
[20]	HERRERA T, NAVARRO DEL HIERRO J, FORNARI T, et al. Inhibitory effect of quinoa and fenugreek extracts on pancreatic lipase and α-amylase under in vitro traditional conditions or intestinal simulated conditions[J]. Food Chemistry,2019,270:509−517. doi: 10.1016/j.foodchem.2018.07.145
[21]	谢晋祥, 耿树香, 宁德鲁, 等. 云南3个主栽品种核桃多肽的制备及体外抗氧化活性的研究[J]. 中国油脂,2025,50(2):51−56,94. [XIE J X, GENG S X, NING D L, et al. Preparation and in vitro antioxidant activity of antioxidant peptides from three main walnut varieties grown in Yunnan[J]. China Oils and Fats,2025,50(2):51−56,94.] XIE J X, GENG S X, NING D L, et al. Preparation and in vitro antioxidant activity of antioxidant peptides from three main walnut varieties grown in Yunnan[J]. China Oils and Fats, 2025, 50（2）: 51−56,94.
[22]	江婷, 费博, 庞会娜, 等. 葛根蛋白及其酶解物的体外抗氧化活性和功能特性[J]. 食品研究与开发,2024,45(1):51−59. [JIANG T, FEI B, PANG H N, et al. Antioxidant activities in vitro and functional properties of Pueraria lobata protein and its enzymatic hydrolysates[J]. Food Research and Development,2024,45(1):51−59.] JIANG T, FEI B, PANG H N, et al. Antioxidant activities in vitro and functional properties of Pueraria lobata protein and its enzymatic hydrolysates[J]. Food Research and Development, 2024, 45（1）: 51−59.
[23]	林娈, 柳雯郡, 黄俊媛, 等. 蛋白核小球藻胰脂肪酶抑制肽的分离纯化、鉴定及其降脂活性[J]. 食品科学,2023,44(24):155−163. [LIN L, LIU W J, HUANG J Y, et al. Isolation, purification, identification and hypolipidemic activity of lipase inhibitory peptide from Chlorella pyrenoidosa[J]. Food Science,2023,44(24):155−163.] LIN L, LIU W J, HUANG J Y, et al. Isolation, purification, identification and hypolipidemic activity of lipase inhibitory peptide from Chlorella pyrenoidosa[J]. Food Science, 2023, 44（24）: 155−163.
[24]	刘晓静. 亚麻籽肽降胆固醇作用的研究[D]. 呼和浩特:内蒙古农业大学, 2020. [LIU X J. Study on the cholesterol lowering effect of flaxseed peptide[D]. Hohhot:Inner Mongolia Agricultural University, 2020.] LIU X J. Study on the cholesterol lowering effect of flaxseed peptide[D]. Hohhot: Inner Mongolia Agricultural University, 2020.
[25]	MCCLEMENTS D J, LI Y. Review of in vitro digestion models for rapid screening of emulsion-based systems[J]. Food Function,2010,1(1):32−59. doi: 10.1039/c0fo00111b
[26]	SHALINI J, SUNDARAPANDIAN T, PRETTINA L, et al. New insights in the activation of human cholesterol esterase to design potent anti-cholesterol drugs[J]. Molecular diversity,2014,18(1):119−31. doi: 10.1007/s11030-013-9464-8
[27]	AWOSIKA O T, ALUKO E R. Inhibition of the in vitro activities of α-amylase, α-glucosidase and pancreatic lipase by yellow field pea (Pisum sativum L.) protein hydrolysates[J]. International Journal of Food Science Technology,2019,54(6):2021−2034. doi: 10.1111/ijfs.14087
[28]	李汉琪, 王治军, 郑清瑶, 等. 多肽组学联合分子对接筛选玉足海参抗血栓肽[J]. 食品与发酵工业,2024,50(20):104−112. [LI H Q, WANG Z J, ZHENG Q Y, et al. Peptidomics combined with molecular docking screening for antithrombotic peptides from sea cucumber (Holothuria leucospilota)[J]. Food and Fermentation Industries,2024,50(20):104−112.] LI H Q, WANG Z J, ZHENG Q Y, et al. Peptidomics combined with molecular docking screening for antithrombotic peptides from sea cucumber （Holothuria leucospilota）[J]. Food and Fermentation Industries, 2024, 50（20）: 104−112.
[29]	FEYISOLA A F, PRITI M, CHEE-YUEN G, et al. Identification and characterization of cholesterol esterase and lipase inhibitory peptides from amaranth protein hydrolysates[J]. Food Chemistry:X,2021,12:100165.
[30]	XIANG H, WATERHOUSE DS, LIU P, et al. Pancreatic lipase-inhibiting protein hydrolysate and peptides from seabuckthorn seed meal:Preparation optimization and inhibitory mechanism[J]. LWT,2020,134:109870. doi: 10.1016/j.lwt.2020.109870
[31]	EGLOFF M P, MARGUET F, BUONO G, et al. The 2.46Å resolution structure of the pancreatic lipase-colipase complex inhibited by a cll alkyl phosphonate[J]. Biochemistry,1995,34(9):2751−2762. doi: 10.1021/bi00009a003
[32]	WANG X, WANG C S, TANG J, et al. The crystal structure of bovine bile salt activated lipase:Insights into the bile salt activation mechanism[J]. Structure,1997,5(9):1209−1218. doi: 10.1016/S0969-2126(97)00271-2
[33]	DANKWA B, BRONI E, ENNINFUL K S, et al. Consensus docking and MM-PBSA computations identify putative furin protease inhibitors for developing potential therapeutics against COVID-19[J]. Structural Chemistry,2022,33(6):2221−2241. doi: 10.1007/s11224-022-02056-1
[34]	TRABUCO L G, LISE S, PETSALAKI E, et al. PepSite:Prediction of peptide-binding sites from protein surfaces[J]. Nucleic Acids Research,2012,40:W423−W427. doi: 10.1093/nar/gks398
[35]	KETPRAYOON T, NOITANG S, SANGTANOO P, et al. An in vitro study of lipase inhibitory peptides obtained from de-oiled rice bran[J]. RSC Advances,2021,11(31):18915−18929. doi: 10.1039/D1RA01411K
[36]	WANG X, AI X, ZHU Z, et al. Pancreatic lipase inhibitory effects of peptides derived from sesame proteins:In silico and in vitro analyses[J]. International Journal of Biological Macromolecules,2022,222:1531−1537. doi: 10.1016/j.ijbiomac.2022.09.259
[37]	YE H, XU Y, SUN Y, et al. Purification, identification and hypolipidemic activities of three novel hypolipidemic peptides from tea protein[J]. Food Research International,2023,165:112450. doi: 10.1016/j.foodres.2022.112450
[38]	GARZÓN A G, CIAN R E, AQUINO M E, et al. Isolation and identification of cholesterol esterase and pancreatic lipase inhibitory peptides from brewer’s spent grain by consecutive chromatography and mass spectrometry[J]. Food and Function,2020,11(6):4994−5003. doi: 10.1039/D0FO00880J
[39]	BABA W N, MUDGIL P, BABY B, et al. New insights into the cholesterol esterase- and lipase-inhibiting potential of bioactive peptides from camel whey hydrolysates:Identification, characterization, and molecular interaction[J]. Journal of Dairy Science,2021,104(7):7393−7405. doi: 10.3168/jds.2020-19868
[40]	NGOH Y Y, CHOI S B, GAN C Y. The potential roles of Pinto bean (Phaseolus vulgaris cv. Pinto) bioactive peptides in regulating physiological functions:Protease activating, lipase inhibiting and bile acid binding activities[J]. Journal of Functional Foods,2017,33:67−75. doi: 10.1016/j.jff.2017.03.029
[41]	NAGAOKA S. Structure-function properties of hypolipidemic peptides[J]. Journal of Food Biochemistry,2019,43(1):e12539. doi: 10.1111/jfbc.12539
[42]	EYDOUX C, SPINELLI S, DAVIS T L, et al. Structure of human pancreatic lipase-related protein 2 with the lid in an open conformation[J]. Biochemistry,2008,47(36):9553−9564. doi: 10.1021/bi8005576